Canrightiopsis, a New Early Cretaceous Fossil with Clavatipollenites-Type Pollen Bridge the Gap Between Extinct Canrightia and Extant Chloranthaceae

Grana

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Canrightiopsis, a new Early Cretaceous fossil with Clavatipollenites-type pollen bridge the gap between extinct Canrightia and extant Chloranthaceae

Else Marie Friis, Guido W. Grimm, Mário Miguel Mendes & Kaj Raunsgaard Pedersen

To cite this article: Else Marie Friis, Guido W. Grimm, Mário Miguel Mendes & Kaj Raunsgaard Pedersen (2015) Canrightiopsis, a new Early Cretaceous fossil with Clavatipollenites-type pollen bridge the gap between extinct Canrightia and extant Chloranthaceae, Grana, 54:3, 184-212, DOI: 10.1080/00173134.2015.1060750

To link to this article: http://dx.doi.org/10.1080/00173134.2015.1060750

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Download by: [Statsbiblioteket Tidsskriftafdeling] Date: 18 January 2016, At: 04:18 Grana, 2015 Vol. 54, No. 3, 184–212, http://dx.doi.org/10.1080/00173134.2015.1060750

Canrightiopsis, a new Early Cretaceous fossil with Clavatipollenites- type pollen bridge the gap between extinct Canrightia and extant Chloranthaceae

ELSE MARIE FRIIS1, GUIDO W. GRIMM2, MÁRIO MIGUEL MENDES3 & KAJ RAUNSGAARD PEDERSEN4

1Department of Palaeobiology, Swedish Museum of Natural History, Stockholm, Sweden, 2Department of Palaeontology, University of Vienna, Vienna, Austria, 3CIMA – Centre for Marine and Environmental Research, University of Algarve, Faro, Portugal, 4Department of Geoscience, University of Aarhus, Aarhus, Denmark

Abstract Canrightiopsis with three species (C. intermedia, C. crassitesta, C. dinisii) is described from the Early Cretaceous of Portugal based on small, one-seeded berries. The fruits are derived from bisexual flowers with three stamens borne on one side of the ovary. There are no traces of a perianth. Pollen is of the Clavatipollenites-type, monocolpate, semitectate, reticulate- columellate with heterobrochate reticulum and muri with beaded supratectal ornamentation. The ovary is unilocular with a single pendant, orthotropous and bitegmic ovule. The seed is endotestal. The endotesta consists of one layer of palisade- shaped crystal cells with fibrous infillings. The fruit wall has resin bodies or cavities from presumed ethereal oil cells sometimes seen as stomata-like structures on the fruit surface. A phylogenetic analysis resolves Canrightiopsis as a close relative of extant Chloranthaceae, particularly close to extant Chloranthus and Sarcandra. All three taxa share the one-sided position of the stamens on the ovary. An evolutionary sequence from fossil Canrightia to fossil Canrightiopsis and extant Chloranthus and Sarcandra is suggested by loss of perianth, reduction in number of ovules and stamens and displacement of stamens to one side of the ovary. Canrightiopsis also shares several critical features with extant Ascarina including monoaperturate pollen and beaded supratectal ornamentation of the pollen wall.

Keywords: Canrightia, Chloranthaceae, Clavatipollenites, Early Cretaceous, endothelium, fossil ﬂowers, synchrotron radiation X-ray tomographic microscopy (SRXTM)

The Chloranthaceae are a small family usually 1960; Walker & Walker 1984)andAsteropollis placed close to the base of the angiosperm phyloge- R.W.Hedl. et G.Norris (compared to pollen of Hedyos- netic tree (APG 2009). The family includes four mum;Walker&Walker1984) occur very early in the extant genera, Ascarina J.R.Forst. et G.Forst., Chlor- history of angiosperms with records extending back to

Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 anthus Sw., Hedyosmum Sw. and Sarcandra Gardner the Barremian (e.g. Couper 1958;Hedlund&Norris that today occur in warm temperate to subtropical 1968; Walker & Walker 1984;Hughes1994; Nichols and tropical regions (Endress 1987). The family has et al. 2006; Friis et al. 2011; Martínez et al. 2013). In a long geological history with an almost worldwide addition to numerous dispersed pollen grains, the fos- distribution in the Cretaceous, and the present dis- sil record of Chloranthaceae also includes a number of tribution and diversity of the family are clearly relic- Cretaceous ﬂoral structures. From the Early Creta- tual. Dispersed pollen grains of Clavatipollenites ceous Hedyosmum-like pistillate and staminate struc- Couper (compared to pollen of Ascarina;Couper tures with Asteropollis-type pollen attached to the ovary

Correspondence: Else Marie Friis, Department of Palaeobiology, Swedish Museum of Natural History, Box 50007, SE-104 05 Stockholm, Sweden. E-mail: [email protected]

(Received 8 March 2015; accepted 19 May 2015)

© 2015 The Author(s). Published by Taylor & Francis. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creative commons.org/licenses/by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way. Canrightiopsis, a new Early Cretaceous fossil 185

or in situ in the stamens have been identified from ovule per ovary as in extant Chloranthaceae. Portugal and eastern North America (Friis et al. Further, Canrightiopsis is similar to Chloranthus and 1997, 2011). From the Late Cretaceous of Scania Sarcandra in having the stamens borne in a partly and eastern North America, inflorescences, isolated epigynous position. Particularly the presence of flowers, androecia and fruits closely resembling those three separate stamen scars in Canightiopsis suggests of extant Chloranthus were assigned to three species a derivation of the tripartite androecium in Chlor- of the extinct genus Chloranthistemon P.R.Crane, anthus from three stamens, not from one. Monocol- E.M.Friis et K.R.Pedersen (Crane et al. 1989;Here- pate, reticulate Clavatipollenites-type pollen similar ndeen et al. 1993;Eklundetal.1997). Chloranthiste- particularly to pollen of extant Ascarina was obser- mon-like floral structures are also known from the mid- ved on the stigmatic surfaces or on the ovary wall of Cretaceous of Germany (Friis et al. 2011; Hartkopf- all three Canrightiopsis species and corroborates pre- Fröder et al. 2011). vious suggestions of a chloranthaceous affinity for Floral structures in all four extant genera of Chlor- some of the early dispersed angiosperm pollen. anthaceae are simple (Endress 1987). Never- theless, the interpretation of certain floral features and basic organisation has not been straightforward. Material and methods fl This to some extent may re ect extensive extinction Material within the lineage and missing data from the geological record. Particularly the nature of the unusual The fossil material reported here was collected from tripartite androecium of Chloranthus has been much several localities (Arazede, Buarcos, Catefica, Juncal- debated (Swamy 1953; Endress 1987). In Chloran- Chicalhão, Famalicão, Vale de Água, Vila Verde) in thus, the androecium is typically three-lobed with a the Lusitanian Basin, western Portugal (for informa- median, tetrasporangiate lobe and two lateral, bis- tion on the mesofossil floras see Friis et al. 2011; porangiate lobes (Endress 1987). Endress (1987) Mendes et al. 2014a). offered two hypotheses for the evolutionary origin Based on stratigraphic information for the Lusi- of the androecium, both starting from a single sta- tanian Basin (e.g. Dinis et al. 2008), the plant bear- men with the three lobes formed either by polymer- ing sediments at the Chicalhão site (Juncal) and the isation, concrescence and reduction, or by lobation Vale de Água locality studied here belong to the and fractionation of a single stamen. basal part of the Famalicão Member of the Figueira New fossil fruits from the Early Cretaceous meso- da Foz Formation established by Dinis (2001). fossil floras of Portugal may fill in some of the miss- Buarcos, Vila Verde and Arazede belong to the ing data critical for understanding the evolutionary basal part of the Calvaria Member of the Figueira steps resulting in the tripartite androecium of extant da Foz Formation established by Dinis (2001). The Chloranthus and fossil Chloranthistemon as well as age of the Famalicão and Calvaria members was other floral features in extant Chloranthaceae. Can- indicated as late Aptian–early Albian by Rey et al. rightia resinifera E.M.Friis et K.R.Pedersen com- (2006). The mesofossils from the Famalicão locality prises small berries with two to five seeds. The were collected in a stratigraphic position below the fruits are derived from bisexual flowers with a partly Calvaria Member indicating a late Aptian age (or epigynous, strongly reduced perianth and four or five older) for this flora. According to Jacques Rey (per- stamens in a radial arrangement on the rim of the sonal communication, June 2012), the plant bearing hypanthium. The seeds are orthotropous, bitegmic sediments of the Catefica locality may belong to the Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 and endotestal very similar to those of Ascarina, younger part of the Almargem Formation that is of Chloranthus and Sarcandra. There are, however, sev- late Aptian–early Albian age. However, the Catefica eral features in Canrightia E.M.Friis et K.R.Pedersen mesofossil flora shares many elements with the late that are not compatible with the extant genera, such Barremian–early Aptian Torres Vedras, also from the as the higher number of ovules per ovary, the pre- Lusitanian Basin, flora and may be slightly older sence of a hypanthium and the radially arranged than the other mesofossil floras studied here, perhaps stamens. belonging to the lower part of the Almargem Forma- Canrightiopsis is a new genus described here that tion (late Barremian–Aptian). shares several features with extinct Canrightia, but that also shares critical features with extant Preparation and analyses of the fossil material Ascarina, Chloranthus and Sarcandra not present in Canrightia. Thus, Canrightiopsis lacks a hypanthium, The fossil plant remains were extracted from the but has stamens (represented by scars) borne on one sediments by sieving in water followed by treatment side of the ovary as is characteristic for extant Chlor- in hydrofluoric and hydrochloric acid, rinsing in anthus and Sarcandra. Canrightiopsis also has a single water and finally air dried. Measurements of fossils 186 E. M. Friis et al.

were made using dissecting microscope as well as of Natural History, Stockholm, Sweden (S) and in from images from scanning electron microscopy the palaeontological collections of the Geological (SEM) and synchrotron radiation-based X-ray Museum of Lisbon, Portugal (P). tomographic microscopy (SRXTM). Pollen grains were measured using SEM images. Phylogenetic analysis For SEM, fossils were mounted on aluminium stubs using nail polish, sputter coated with gold To explore the systematic affinity of the new fossils and studied using a Phillips 515 Scanning Electron from Portugal, they were included in the morpholo- Microscope at 15 kV, and a Hitachi S-4300 Field gical matrix of Doyle and Endress (2010; D&E matrix Emission Scanning Electron Microscope at 2 kV. in the following). We also included the closely similar Internal structures were studied using SRXTM at fossils Canrightia resinifera and Zlatkocarpus J.Kvaček the TOMCAT beamline of the Swiss Light Source at et E.M.Friis in the analysis following Doyle and End- the Paul Scherrer Institute, Villigen, Switzerland ress (2014). A reduced morphological matrix includ- (Stampanoni et al. 2006). About 30 specimens ing only characters that were observed for at least one were mounted on brass stubs with a diameter of of the three fossil taxa (leaving 63 out of a total of 142 3 mm using nail polish. All specimens were mea- characters) was analysed using PAUP* (Swofford sured at 10 keV using a sCMOS detector and a 2002) to calculate pairwise mean morphological dis- 20 μm thick LAG:Ce scintillator screen. We used a tances. SPLITSTREE 4 (Huson & Bryant 2006) was 10× objective with isotropic pixel size of 0.65 μm then used on the distance matrix to infer a neighbour- (P0311, S174006, S174027, S174028, S174039, net splits graph (NNet; Bryant & Moulton 2002, S174040, S174155, S174157, S174159, S174174, 2004). The NNet’s ability to express signal incompat- S174248, S174309, S174310, S174311) or 20× ibility in an underlying matrix has proven to be useful objective with isotropic pixel size of 0.325 μm in phylogenetic assessment of fossil taxa to overcome (S174023–S174026, S174033, S174104, S174105, problems with homoplasy as found also in the D&E S174107, S174148–S174153, S174175). A more matrix (Denk & Grimm 2009; Friis et al. 2009; detailed outline of the technique used for coalified Mendes et al. 2014b). plant fossils is given by Friis et al. (2014). Slice data derived from the scans (Hintermüller et al. 2010) were analysed and manipulated using AVIZO (v6.1 Character plot and v7.1) software for computed tomography. Theevolutionarysignificance of character traits All specimens and preparations of the new taxon expressed by Canrightiopsis was evaluated by map- from Portugal described here are housed in the ping 13 morphological characters on a newly gen- palaeobotanical collections of the Swedish Museum erated molecular-based tree. The selected charac-

Table I. Selected morphological characters for Canrightia, Canrightiopsis, Zlatkocarpus and extant Chloranthaceae.

No. Characters Hedyosmum Ascarina Chloranthus Sarcandra Canrightiopsis Canrightia Zlatkocarpus

1 Bisexual/unisexual- 21000 0 ? pseudobisexual/unisexual 2 Tepals present /absent 0 1 1 1 1 0 0

Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 3 Partly epigynous org./ 1?000 00 epigynous org. 4 Stamens more than one/one 1 0/1 0 1 0 0 ? 5 Distal aperture regular/distal 10220 00 aperture irregular/distal aperture absent 6 Supratectal spines absent/ 1 1 0/1 0 1 0 0 present 7 Carpels one/2–5000001? 8 Style absent/present 1 0 0 0 0 0 0 9 Oil cells intrusive/internal/ 21000 0 ? absent 10 Fruit ﬂeshy/dry 1 0 0 0 0 0 0 11 Endotesta unspecialised/ 01111 1 ? endoreticulate 12 Integuments tanniferous/not 1 0 0/1 1 0 0 ? tanniferous 13 Distinct endotelium yes/no 1 1 1 1 1 0 ? Canrightiopsis, a new Early Cretaceous fossil 187 Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016

Figure 1. SEM images of Canrightiopsis intermedia gen. et sp. nov. Fruits, from the Early Cretaceous Famalicão locality, Portugal; holotype (S174033; sample Famalicão 25). A–F. Fruit rotated to document the position on the three stamen scars (numbered 1–3) on one side of the fruit (dorsal), stamen 2 is central, 1 and 3 in more lateral positions on the dorsal side; there are no stamen scars on the other (ventral) side; note scattered openings in the epidermis of the fruit wall representing ethereal oil cells. G–I. Magniﬁcation of the three dorsal stamen scars; numbering the same as for (A–F). Scale bars – 500 µm (A–F), 100 µm (G–I).

ters are traits that can be observed in the fossil per 24 July 2014 were harvested from gene banks material and that differ between extant members using the same protocol and software environment of the Chloranthaceae. All available sequence data as in Grimm and Renner (2013). Information in 188 E. M. Friis et al.

&Maddison2011) was then used to reconstruct possible ancestral states for the 13 selected, variable characters along the molecular based topology under maximum parsimony (MP) with the standard parsimony model where all characters are treated as unordered and under maximum likelihood (ML) using Lewis’ one-parameter Markov-model (Lewis 2001). The selected morphological characters (Table I) follow Doyle and Endress (2010, 2014) with a few exceptions. Canrightia is scored as having a partly epigynous ovary. We scored Canrightiopsis, Chlor- anthus and Sarcandra in a similar way based on the position of the stamens and stamen scars on the ovary, about one third to two thirds up. We also scored Chloranthus as having more than one stamen based on its full stamen and the additional lateral half stamens. We follow Doyle and Endress (2014) in included the extinct Zlatkocarpus in the analysis, but changed some of the scoring for Zlatkocarpus to express the uncertainty in the interpretation of this fossil (sexuality here scored as unknown in contrast to unisexual in Doyle & Endress 2014; aperture membrane scored as unknown in contrast to sculp- tured in Doyle & Endress 2014).

Results Small elliptical to ovate berries occur abundantly in the Early Cretaceous plant bearing strata of Portu- gal. We here describe some of these berries characterised by the presence of stamen scars borne on one side of the fruit wall. There are no traces of a Figure 2. SEM images of Canrightiopsis intermedia gen. et sp. nov. perianth or hypanthium. Each fruit has a single Fruits, from the Early Cretaceous Famalicão locality, Portugal; (S174107; sample Famalicão 25). A, B. Fruit in apical (A) and orthotropous and distinctly foveolate, endotestal dorsal (B) view showing the position on the three stamen scars seed. The fruits resemble in some aspects fossil (numbered 1–3), position of stigmatic area indicated by asterisk. berries of the extinct Canrightia also described Scale bars – 500 µm. from the Early Cretaceous of Portugal (Friis & Ped- ersen 2011), but they are distinct in a number of external and internal traits. We include these single- NCBI GenBank flatfiles were read out using seeded berries in Canrightiopsis gen. nov. Currently, Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 GBK2FAS (Göker et al. 2009), alignment relied almost 1000 specimens of isolated fruits and iso- on MAFFT v7 (Katoh & Standley 2013), strict- lated seeds of Canrightiopsis have been recovered genus consensus sequences were generated with from the sediments. Most are from the Famalicão G2CEF (Göker & Grimm 2008). RAXML v7.2.8 locality, where about 650 specimens have been was used to find a topology with high likelihood separated from the organic residue so far. Within and establish branch support via quick, non-para- the collection, there is variation in size of fruits, metric bootstrapping using a sufficient number of seeds and pollen as well as details of the seed wall bootstrap replicates (Stamatakis 2006; Stamatakis suggesting the presence of several species. Based on et al. 2008;Pattengaleetal.2009). The final matrix our current knowledge three species, Canrightiopsis comprised 16 779 characters (nucleotide sites) for intermedia sp. nov., C. crassitesta sp.nov.andC. the five extant genera of Chloranthaceae, covering dinisii sp. nov., are defined. In many specimens, two nuclear (18S rDNA, 25S rDNA), three mito- the fruit wall is abraded or strongly wrinkled and chondrial (atp1, matR, nad5) and five plastid gene the scars, interpreted as stamen scars (see Descrip- regions (atpB, matK, ndhF, rbcL, rpoC2) (see also tion of the fossils and Discussion sections), are Supplemental data). MESQUITE v2.75 (Maddison often difficult to detect and these specimens are Canrightiopsis, a new Early Cretaceous fossil 189

unassigned at the specific level. Generic and speci- pendant, orthotropous ovules/seeds with a distinct ficdiagnosesaregiveninthesystematicsection endotesta composed of a single layer of palisade- followed by a collective description of all Canright- shaped crystal cells with fibrous infilling. In both iopsis fossils with references to species specific Canrightiopsis and Canrightia, stamen scars are features. placed about one to two thirds up the ovary/fruit wall. Both taxa have semitectate-reticulate and monocolpate pollen. Canrightiopsis differs, however, Systematics from Canrightia in having single-seeded berries in Angiospermae contrast to the two- to five-seeded berries of Can- Chloranthaceae rightia. Only in one case, a two-seeded berry has been observed for a specimen that may belong to Canrightiopsis gen. nov. Canrightiopsis, while no one-seeded berry is known Derivation of generic name. — From opsis (Greek: for Canrightia. Stamen scars in Canrightiopsis are not appearance) to indicate close similarity in morpho- subtended by perianth parts and there is no logical appearance to fossil fruits of Canrightia. hypanthium as there is in Canrightia. Further, in Canrightiopsis stamen scars are restricted to one side Diagnosis. — Flower bisexual. Perianth lacking. of the ovary, while stamen scars in Canrightia are Androecium of three stamens. Stamens restricted to radially arranged. Pollen of all Canrightiopsis species one side of the ovary, borne about one third to two is of the Clavatipollenites-type with beaded muri, thirds up the ovary. Stamens leaving distinct scars while pollen of Canrightia is of the broadly defined on the surface of the fruits. Associated pollen of Cla- Retimonocolpites-type with smooth muri. Seeds of vatipollenites-type; pollen monocolpate with long, Canrightiopsis are also distinguished from those of extended aperture; pollen wall semitectate, reticu- Canrightia by their much thicker endotesta (50– late-columellate; reticulum heterobrochate; columel- 120 µm in Canrightiopsis; about 20 µm in Canrightia). lae long, irregularly distributed; colpus membrane Crystals of the endotesta are irregularly distributed verrucate-rugulate; colpus margin indistinct. Ovary in Canrightiopsis with most crystals towards the mid- semi-inferior inferred from the position of the stamen dle and inside of the endotesta while in Canrightia, scars; unilocular with a single pendant ovule. Stigma large crystals are densely packed in the outer part of not or only slightly raised, indistinct or hemispherical. the endotesta. Ovule orthotropous, bitegmic, endotestal; endotesta Canrightiopsis is distinguished from fruits, seeds thick with one layer of palisade-shaped crystal cells and pollen of the Early and mid-Cretaceous Couper- with fibrous infillings; single layer of larger, thin- ites K.R.Pedersen, P.R.Crane, Drinnan et E.M.Friis. walled cells outside endotesta; tegmen of several Both taxa have one-seeded fruits, but seeds of Cou- layers of thin-walled cells. Fruit single-seeded berry. perites are anatropous and exotestal and the pollen Fruit wall with resin bodies or cavities from presumed have a distinct, sharp colpus margin (see Discussion ethereal oil cells that on the outer surface of fruit wall section). are seen as stomata-like structures. Embryo in mature The fossil fruits and seeds of Canrightiopsis also seeds tiny. share many features with fruits and seeds of extant Ascarina, Sarcandra and Chloranthus, but are distin- Type species designated here. — Canrightiopsis interme- guished particularly based on androecial features (no dia sp. nov. stamen on the ovary/fruit wall of Ascarina; only one Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 stamen in Sarcandra; tripartite androecium in Comments on the genus. — The fruits and seeds of Chloranthus). Canrightiopsis are similar to those of the extinct Can- rightia in several respects. Both genera have small Canrightiopsis intermedia sp. nov. berries with resin bodies in the fruit wall and Figures 1–4, 7, 10, 11

→ Figure 3. SEM images of Canrightiopsis intermedia gen. et sp. nov. Fruits, from the Early Cretaceous Famalicão locality, Portugal. A. Dorsal view of fruit with three stamen scars (numbered 1–3) on a slightly raised rim; position of stigma marked by asterisk (S174018; sample Famalicão 25). B. Dorsal view of fruit with three stamen scars (numbered 1–3) and a bract (b) attached to the basal part of the fruit (S174156; sample Famalicão 25). C. Fruit in partly ventral view with many openings in the epidermis of the fruit wall showing distribution of ethereal oil cells (S174004; sample Famalicão 25). D. Detail of fruit wall showing epidermal cells with wrinkled cuticle and stomata-like openings (S107700; sample Famalicão 25). E. Detail of fruit surface of holotype showing stomata-like openings (S174033; sample Famalicão 25). F, G. Details of fruit in ﬁgure (C) showing remains of resin in an abraded oil cell (F) and intact stomata-like opening over oil cell (G) (S174004; sample Famalicão 25). Scale bars – 500 µm (A–C), 100 µm (D–F), 20 µm (G). 190 E. M. Friis et al. Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 Canrightiopsis, a new Early Cretaceous fossil 191 Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 192 E. M. Friis et al.

Derivation of specific epithet. — From inter and med- Derivation of speci fic epithet. — From crassus (Latin: ium (Latin: between and middle) to indicate an thick, solid) for the thick endotesta. intermediate form between fossil Canrightia and extant Chloranthus. Specific diagnosis. — As for the genus with the following addition: Fruit elliptical to ovate. Endotesta Specific diagnosis. — As for the genus with the fol- thick. Stigmatic area only slightly raised. lowing addition: Fruit elliptical to ovate. Endotesta moderately thick. Stigmatic area only slightly raised. Dimensions. — Fruits 0.85–1.05 mm long and 0.6– 0.8 mm broad. Pollen 10.3–12.4 µm in equatorial diameter. Endotestal cells about 80–120 µm high. Dimensions. — Fruits 1.1–1.3 mm long and 0.9– 1.15 mm broad. Pollen 13–14.5 µm in equatorial Holotype designated here. — S174311 (sample diameter. Endotestal cells about 50–90 µm high. Catefica 343); illustrated here in Figures 5A,D– F, 12A–C. Holotype designated here. — S174033 (sample Fama- – – licão 25); illustrated here in Figures 1A I, 11A C. Paratypes designated here. — S174039, S174159 (sample Catefica 49), S174310 (sample Catefica Paratypes designated here. — S107700–S107708, 154); S122089 (sample Catefica 342). S174004–S174006, S174016–S174028; S174107, S174108, S174148, S174150–S174158, S174364– Type locality. — Catefica, Portugal (39° 03ʹ 30ʺ N; S174368 (sample Famalicão 25). Totally, about 50 09°14ʹ 30ʺ W). specimens, several specimens stored in slide boxes under the same number. Type horizon and age. — Early Cretaceous (late Bar- remian–early Albian; Almargem Formation). Other specimens. — S 174104, S174105 (sample Buarcos 157); S174174 (sample Vale de Água 331). Distinguishing features for Canrightiopsis crassitesta. — Canrightiopsis crassitesta is distinguished from C. Type locality. — Famalicão, Portugal (39° 42ʹ 16ʺ intermedia by its thicker endotesta and from C. dinisii N; 08° 46’ 12ʺ W). by its more elliptical to ovate fruits and generally thicker endotesta. Type horizon and age. — Early Cretaceous (Aptian; Canrightiopsis dinisii sp. nov. below the Calvaria Member of the Figueira da Foz Figures 6, 9, 13 Formation). Derivation of specific epithet. — In honour of Jorge Distinguishing features for Canrightiopsis intermedia. — Manuel Leitão Dinis for his contribution to the Creta- Canrightiopsis intermedia is distinguished from C. cras- ceous geology of Portugal. sitesta and C. dinisii mainly by its thinner endotesta. Specific diagnosis. — As for the genus with the fol- Canrightiopsis crassitesta sp. nov. lowing addition: Fruit almost spherical. Endotesta

Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 Figures 5, 8, 12. thick, stigmatic area distinct.

← Figure 4. SEM (A–C, H) and SRXTM images (D–G, I)ofCanrightiopsis intermedia gen. et sp. nov. Fruits, from the Early Cretaceous Buarcos locality, Portugal. A, B Fruit in two different views (A: dorsal to lateral view, B: dorsal view) showing three stamens (numbered 1–3) on the dorsal side of the fruit; surface of fruit with scattered holes representing the ethereal oil cells; position of stigma marked by asterisk (S174105; sample Buarcos 157). C. Detail of fruit in (A, B) showing magnification of the central stamen scar. D, E. Volume rendering (D) and orthoslice (E) of fruit in (A, B) cut longitudinally through middle of fruit exposing enclosed seed with a thick endotesta (o-en) and thin tegmen (i-te); holes are present in the outer part of endotesta; vascular supply (arrowhead) to chalaza (ch) enters through narrow opening in endotesta; micropylar area of tegmen pointed (arrow, mi); remnants of protruding stamen scar seen on dorsal side (2). F. Tangential and longitudinal orthoslice through endotesta of same specimen as shown in (A–E) showing irregular holes arranged in longitudinal grooves. G. Detail of same specimen showing the high palisade cells of endotesta (o-en) with fibrous infilling. H. Abraded fruit exposing enclosed seed with foveolate surface of the endotesta (S174104; sample Buarcos 157). I. Transverse orthoslice through specimen in (H) showing collapsed cells of fruit wall enclosing distinct endotesta with scattered holes in the outer surface and remains of endosperm in the centre. Scale bars – 500 µm (A, B, D, E, H, I), 200 µm (F, G), 50 µm (C). Canrightiopsis, a new Early Cretaceous fossil 193

Figure 5. SEM (A–C) and SRXTM images (volume renderings D–F)ofCanrightiopsis crassitesta gen. et sp. nov. Fruits, from the Early Cretaceous Cateﬁca locality, Portugal. A, D. Holotype, view showing two of three stamen scars (numbered 2–3) borne on a slightly raised

Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 rim and wrinkled surface of fruit wall (S174311; sample Cateﬁca 343). B. Strongly compressed and abraded fruit with enclosed foveolate seed exposed (S122089; sample Cateﬁca 342). C. Detail of holotype showing epidermal cells with stomata-like structures representing ethereal oil cells and wrinkled cuticle. E, F. Volume renderings of holotype cut longitudinally (E: tangential, F: through middle of seed) showing foveolate surface of endotesta (E) and thick endotesta and vascular supply for chalaza (F, arrowhead). Scale bars – 500 µm (A, B, D–F), 100 µm (C).

Dimensions. — Fruits 1.1 mm long and 0.9 mm Other specimens. — P0095, P0096 (sample Chical- broad. Pollen 14 µm in equatorial diameter. Endo- hão 96), P0313, P0314 (sample Chicalhão 127); testal cells about 100 µm high. P0315, P0316, P0317, P00318, P0319, P0320, P0321, P0322, P0323, P0324, P0325, P0326 (sam- Holotype designated here. — P0311 (sample Chical- ple Chicalhão128). hão 125); illustrated here in Figures 6A, B, F, 9A–C, 13A–D. Type locality. — Chicalhão opencast clay pit com- plex near Juncal, Portugal (39° 35ʹ 34.8ʺ N08°54ʹ Paratypes designated here. — P0312 (sample Chical- 19.2ʺ W). hão 125). 194 E. M. Friis et al. Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016

Figure 6. SEM images of Canrightiopsis dinisii gen. et sp. nov. Fruits, from the Early Cretaceous Chicalhão site, Portugal (sample Chicalhão 125). A. Fruit with two of three stamen scars exposed (numbered 1–2) and slightly raised, rounded stigmatic area (asterisk) (holotype, P0311). B. Detail of holotype showing stamen scar (1). C. Slightly abraded fruit with two of three stamen scars (numbered 1 and 3) exposed; a further, larger scar-like structure (arrowhead) is different from the stamen scars in other specimens; stigmatic area (asterisk) indistinct (P0312). D. Detail of larger scar of fruit shown in (C) probably representing oil cell opening or damage to the fruit wall. E. Details of fruit shown in (C) showing stigmatic area (asterisk) and epidermal cells of fruit wall. F. Detail of holotype showing epidermal cells of fruit wall with stomata-like openings. Scale bars – 500 µm (A, C), 100 µm (B, D, E), 50 µm (F). Canrightiopsis, a new Early Cretaceous fossil 195 Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016

Figure 7. SEM images of Canrightiopsis intermedia gen. et sp. nov. Stigmatic area and pollen, from the Early Cretaceous Famalicão locality, Portugal (sample Famalicão 25). A. Stigmatic area covered by amorphous substance with pollen grains embedded (S174018). B. Group of monocolpate pollen clustered on fruit surface showing apertures (S174005). C. Pollen grains close to stigmatic area of fruits in (A). D. Pollen grains on surface of fruit in Figure 3C (S174004). E. Pollen grains on surface of fruit showing reticulum, columellae and beaded muri (S107702). F. Pollen grains on surface of fruit with pollen wall folded over aperture; reticulum well exposed showing scattered columellae (S107703). Scale bars – 100 µm (A), 50 µm (B), 10 µm (C–F). 196 E. M. Friis et al. Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016

Figure 8. SEM images of Canrightiopsis crassitesta gen. et sp. nov. Pollen, from the Early Cretaceous Catefica locality, Portugal. A–D. Pollen grains from surface of fruit shown in Figure 5B showing heterobranchiate reticulum and beaded supratectal ornamentation; pollen with wall folded over aperture (A, B); pollen with exposed aperture (C) showing verrucate aperture membrane and indistinct margins; detail of pollen wall showing supratectal ornamentation (D) (S122089; sample Catefica 342). E–G. Pollen grains from surface of holotype (S174311; sample Catefica 343). Scale bars – 10 µm (A), 5 µm (B, C, E–G), 2.5 µm (D). Canrightiopsis, a new Early Cretaceous fossil 197

Type horizon and age. — Early Cretaceous (late and show that the fruits are derived from bisexual Aptian–early Albian, Famalicão Member of the Fig- flowers. A single fruit of Canrightiopsis intermedia ueira da Foz Formation). from Famalicão has one small, fan-shaped subtending bract preserved adhering to the base of the fruit Distinguishing features for Canrightiopsis dinisii. — (Figure 3B) showing that the fruit is developed from Canrightiopsis dinisii is distinguished from C. intermedia an almost sessile flower. This is also supported by and C. crassitesta in its more spherical shape and dis- the morphology of the isolated fruits that lack stalks tinct stigmatic area. It is further distinguished from C. or have remnants of a very short stalk (Figure 4D, intermedia by its much thicker endotesta. E). There is no other information on how the fruits were borne on the plant. The specimen with sub- Canrightiopsis sp. tending bract preserved is important in allowing observations on the orientation and position of the A number of specimens cannot be included with ovule and androecium, although the orientation can- certainty in any of the three species established here not be established with certainty without develop- for Canrightiopsis. Most of them are abraded fruits mental information. or isolated seeds where information on stamen The fruits of Canrightiopsis intermedia and C. cras- scars, pollen or seed wall is missing. We refer to sitesta are typically ovate to elliptical, sometimes flat- these specimens only as Canrightiopsis sp. Canright- tened on one side (Figures 1A–F, 2A, B, 3A–C, 4A, iopsis is most abundant in the Famalicão flora with B, D, H, 5A, B, D, E) while fruits of C. dinisii are about 600 specimens assigned to Canrightiopsis sp.: almost spherical (Figure 6A, B). The surface is typi- S107695–S107699, S107709, S107710, S174149, cally strongly wrinkled and it may be difficult to S174369–S174378 (sample Famalicão 25). Many observe details on the fruit wall. Internal details of them probably belong to C. intermedia.The were studied using SRXTM for 19 specimens from genusisalsocommonintheCatefica mesofossil Famalicão (S174006, S174023–S174028, S174033, flora with about 150 specimens assigned to Can- S174107, S174148–S174153, S174155, S174157, rightiopsis sp.: S174379, S17434380, S174396 S174159, S174175), six specimens from Catefica (sample Catefica 49); S174381 (sample Catefica (S172309, S174039, S174040, S174248, S174310, 50); S174382 (sample Catefica 51); S174383 S174311), two specimens from Buarcos (S174104, (sample Catefica 150); S174384 (sample Catefica S174105), one specimen from Vale de Água (S174174) 152); S174385 (sample Catefica 153); S174309, and one specimen from Chicalhão (P0311). Based on S174386 (sample Catefica 154); S174387 (sample these studies, it was possible to reconstruct morphology Catefica 242); S174388 (sample Catefica 342); of seeds enclosed in the fruits and thus to link the fruits S174389 (sample Catefica 343); S174390 (sample to isolated seeds co-occurring associated with the fruits. Catefica 359); S174391 (sample Catefica 360); There are also a number of specimens where seeds have S174392 (sample Catefica 361); S174393 (sample part of the fruit wall preserved. Catefica 362); S174394 (sample Catefica 381); No stamens are found attached, but in better pre- S174395 (sample Catefica 382). The genus is less served specimens three distinct scars are present common in the other mesofossil floras. Twenty-two about one third to two thirds up on the fruit. The specimens are recorded from the Arazede locality: scars are restricted to one side of the fruit and typi- S174457 (sample Arazede 372), S174458 (sample cally protrude from the fruit surface (Figures 1A–I, Arazede 374); seven specimens from the Buarcos 2A, B, 3A, B, 4A–C, 5A, D, 6A–C). The size and Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 locality: S174459, S174460 (sample Buarcos 157), general appearance of the scars indicate that they are S174461 (sample Buarcos 209); and four speci- from stamens. One specimen of Canrightiopsis inter- mens from the Vila Verde 2 locality: S174463 media (S174156) with a fan-shaped subtending bract (sample Vila Verde 2 439, S174464 (sample Vila still attached to the fruit has the three stamen scars Verde 2 441), S174465 (sample Vila Verde 2 442). on the same side as the subtending bract (Figure 3B) Several specimens are stored in slide boxes under demonstrating that the stamens are probably on the same number. dorsal side of the carpel, although this cannot be established with certainty as developmental sequences are lacking. Because some of the fruits are Description of the fossils strongly wrinkled or partly abraded, the scars are No flowers that can be linked to the fossil fruits have not always clear. In some specimens, there is a dis- been discovered in any of the Early Cretaceous tinct rim on the androecium side of the ovary sup- mesofossil floras, but distinct scars from stamens porting the scars (Figures 3A, B, 5A), but this is not (see later) have been observed in all three species always the case and, except for the occasional rim, (Figures 1A–I, 2A, B, 3A, B, 4A–C, 5A, D, 6A–D) there are no features that could indicate traces of a 198 E. M. Friis et al.

hypanthium. The scars are all of the same dimension an amorphous substance (Figures 7A, B, 9A–C) indicating that they are scars from equally developed suggesting that the stigma was wet. stamens. They are placed with one scar in a central The fruits are one-seeded berries, 0.85–1.3 mm position (no. 2 in Figures 1–6) and one scar on both long and 0.6–1.15 mm broad, with a thin fruit wall sides (no. 1 and no. 3 in Figures 1–6) of the central of small parenchyma cells (Figures 4I, 11B, 13A–C). scar with a distance between the scars of about The soft tissue of the fruit is usually strongly com- 300 µm (Figures 1A–F, 2A, B). Sometimes, there pressed or not preserved and often, the outer cuticle are also other scar-like structures that appear mostly of the fruit wall is collapsed over the hard endotesta to be from burst oil cells, but could also be from of the seed (Figure 10A, B). Scattered on the outer various damages to the fruit wall. In one of the two surface of the fruits, are numerous stomata-like fruits of C. dinisii from the Chicalhão mesofossil structures. In well preserved specimens, they are flora, an additional, prominent scar is present intact with smooth cells in contrast to the other (Figure 6C, D). It is distinguished from the other epidermal cells of the fruit wall that usually show scars in being bigger and not protruding. The nature distinct cuticular ridges or wrinkles (Figures 3D–G, of this scar is uncertain, but the possibility that it 5C, 6E, F). These stomata-like structures are similar represents a scar from an aborted stamen cannot be to the ethereal oils cells seen on the carpel surface of ruled out. Another possibility is that it is a burst oil Chloranthus spicatus (Thunb.) Makino (Endress & cell or damage to the epidermis. Vascular bundles Igersheim 1997) and are most likely also ethereal supplying the stamens are rarely preserved, but can oil cells. The cells are often broken leaving small be seen in a specimen from Vale de Água. holes scattered over the fruit wall (Figures 1A–F, Pollen is observed on the surface of several fruits 3B, C, 4A, B), and in some specimens, these holes or on the stigmatic surface (Figures 7–9). Pollen contain resinous material (Figure 3F) corroborating grains are semitectate-reticulate and monocolpate, the interpretation of the structures as ethereal oil almost circular in equatorial outline and 10–15 µm cells. in diameter (Canrightiopsis intermedia:13–14.5 µm; There is a single orthotropous, bitegmic and endo- C. crassitesta: 10.3–12.4 µm; C. dinisii:14–14.9 µm). testal seed in each fruit (Figures 4D, E, H, I, 11A–D, The reticulum is heterobrochate. Details of the pol- 12A–H, 13A–C). The seed is pendant and ventral len sculpture are well-exposed in some specimens of inferred from the position of the subtending bract in C. intermedia (Figure 7) and specimens of C. crassi- specimen S174156 from Famalicão. A vascular bundle testa (Figure 8), while grains on the stigma surface of extends on the presumed ventral side of the fruit C. dinisii (Figure 9) are embedded in a secretion that (opposite the stamen scars) from the base of the fruit impedes detailed observations. It is, however, clear to the level of the chalaza and stigma (Figure 4D, 5F, that the pollen of all three species of Canrightiopsis is 12D). There are also vascular bundles on the side of the Clavatipollenites-type with beaded muri. Muri bearing the stamen scars, but the fruit wall is usually are narrow with rounded profile and an ornamenta- poorly preserved and the bundles are difficult to trace. tion of low supratectal elements. Columellae are A single fruit from the Catefica locality (S172333) long and irregularly distributed. The colpus mem- diverges from all other fruits assigned to Canrightiopsis brane is covered by irregular verrucate-rugulate ele- in having two seeds. The seed wall is comparable to ments that also have a beaded ornamentation. The that of other Canrightiopsis species in general structure, colpus margin is indistinct and there is a gradual but the specimen is compressed and somewhat frag- transition from the verrucate-rugulate elements of mented and details on seed attachment and scars are Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 the colpus membrane to the open reticulum of the uncertain. intercolpi areas (Figure 8C). The chalaza is indicated by a thickening in the The gynoecium is unilocular, apparently monocar- exotegmen (Figures 4E, 12A, D). The micropyle is pellate with a single subapical and rounded stigma placed opposite of the chalaza and marked by a that is usually only slightly raised and about 100 µm narrow opening in the endotesta and a thickening in diameter. In Canrightiopsis dinisii, it is distinct of the cuticle of the exotegmen (Figure 11D); cells of and hemispherical (Figures 6A, 9A). It is uncertain exotegmen in the micropylar region are sinuous. whether a similar raised stigma was also present in The seed wall is mainly made up by the outer the other species, but has been abraded during fossi- integument (testa). The endotesta is composed of lisation. Considering the numerous specimens pre- one layer of palisade-shaped cells, 50–120 µm high sent in the Famalicão and Catefica mesofossil floras, (Canrightiopsis intermedia: about 50–90 µm; C. crassi- it is, however, most likely that the stigma is indistinct testa: about 80–120 µm; C. dinisii: about 100 µm). in the other species of Canrightiopsis. Pollen is some- So far, seeds with a very thick endotesta have only times attached to the stigma and often embedded in been observed from specimens from the Catefica Canrightiopsis, a new Early Cretaceous fossil 199

mesofossil flora assigned to C. crassitesta, but there is 1.3 mm). They may represent different developmental a considerable variation in endotesta thickness of stages or perhaps other species. Unless other charac- Canrightiopsis in most mesofossil floras. The endo- ters are available such as internal structures obtained testa cells have fibrous infilling and crystals (Figures 4E–G, I, 11A–F, 12A–H, 13A–C). In surface view, the endotestal cells are isodiametric and polygonal (Figure 10F). Crystals are concentrated mainly in the inner and middle part of endotesta. The endotesta is characterised by small holes placed in longitudinal grooves that give the seed surface a ribbed appearance (Figures 4F,H,I,5D, E, 10A–F). A layer of larger, isodiametric cells is preserved outside the endotesta. These cells have thin cell walls and sometimes an infilling of amorphous material that is interpreted as tannin (Figures 11A–C, 13A–C). It is possible that these cells represent the epidermis of the testa (exotesta) and that the testa is only two cell layers thick, but there is not a clear delimitation between this layer and the compressed cells of the fruit wall. Above the holes in the endotesta, there may be additional thin walled cells. The inner integument (tegmen) consists of several layers of thin-walled cells (Figure 12F–H), but these are often collapsed (Figure 11A–C). One specimen has radially elongated cells of the inner integument (Figure 11E, F) that show some resemblance to the endothelium cells observed in Canrightia and extant Lactoris Phil. This specimen is probably aborted or immature. Other specimens show a particular pat- tern of a fragmented tegmen with elongated cells (Figure 13A–C) that may also be remains of an endothelium, but none of the Canrightiopsis specimens shows a distinct endothelium. A schematic overview of the fruit and seed wall is shown in Figure (14). The embryo is small and less than one quarter of the total length of the seed. It is rarely preserved, but may leave a hollow space in the surrounding endosperm in well-preserved, mature seeds (Figure 12A– C). Sometimes, part of the embryo may also be preserved (Figure 11D). Seeds are found inside the fruits as well as isolated, Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 sometimes with fruit wall remnants preserved (Figures 4H, 5B, 10B–E). Isolated seeds, characterised by the foveolate and grooved surface, are more common in the mesofossil flora than the fruits and vary much more in size than the fruits (0.6–1.5 mm long and 0.45–

→ Figure 9. SEM images of Canrightiopsis dinisii gen. et sp. nov. Stig- matic area and pollen, from the Early Cretaceous Chicalhão site, Portugal (holotype P0311; sample Chicalhão 125). A. Raised stigmatic area with numerous pollen grains embedded in amorphous substance. B–C. Details of pollen from stigmatic area showing monocolpate aperture and reticulate tectum (B)aswellassupratec- talornamentation.Scalebars100µm(A),20µm(B),10µm(C). 200 E. M. Friis et al.

Figure 10. SEM (A–E) and SRXTM images (F)ofCanrightiopsis intermedia gen. et sp. nov. Fruits and seeds, from the Early Cretaceous Famalicão locality, Portugal (sample Famalicão 25). A, B. Fruits with fruit wall collapsed over the foveolate surface of the endotesta (A: S107707; B: S174026). C–E. Foveolate seeds with holes in endotesta in longitudinal grooves (C: S174024; D: S174025; E: S174023). F. Longitudinal and tangential section of endotesta (221 consecutive orthoslices) showing holes in endotesta and polygonal and isodiametric facets of palisade cells. Scale bars – 500 µm (A–E), 100 µm (F). Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 from SRXTM analysis, the isolated seeds have been Chloranthaceae (Ascarina, Chloranthus, Sarcandra), treated as Canrightiopsis sp. with Canrightiopsis being very similar to extant Chlor- anthus and Sarcandra in its preserved morphological traits. Character scoring and phylogenetic analysis Character mapping of 13 morphological traits Out of the 142 characters in the D&E matrix, 48 variable within modern Chloranthaceae unequivo- could be scored for the new fossil genus Canrightiopsis cally places Canrightiopsis in the core Chloranthaceae described here compared to 57 defined characters for (Figure 16). The character suite of Canrightiopsis Canrightia and 34 for Zlatkocarpus, providing a total of brings it close to the putative common ancestor of 63 characters defined for at least one of the three fossil Chloranthus and Sarcandra, independent of the root of taxa. This reduced character set is sufficient to group Chloranthaceae, either with Hedyosmum as sister to the all extant Chloranthaceae except for Hedyosmum rest, as inferred from most molecular data, or with in the resultant neighbour-net (Figure 15). The Hedyosmum as sister to Ascarina, as traditionally per- three fossil taxa, Canrightia, Canrightiopsis and Zlatko- ceived based on morphological evidence, such as pre- carpus were resolved as close relatives of the core sence of unisexual flowers and monoaperturate pollen. Canrightiopsis, a new Early Cretaceous fossil 201 Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 202 E. M. Friis et al.

Discussion Only mature fruits of Canrightiopsis have been recovered, but based on the distinct stamen scars on The small one-seeded berries and isolated, foveolate the fruit wall, we conclude that the Canrightiopsis seeds assigned here to Canrightiopsis gen. nov. occur flowers were bisexual with an androecium of three abundantly in the Early Cretaceous plant-bearing stamens in a one-sided position on the ovary. All sta- sequence of western Portugal. Judged from the abun- mens are shed, but the three scars on the fruits are of dance of specimens and the widespread occurrence in similar size and shape indicating that all three stamens the mesofossil floras, extending from Catefica in the of the flower were equally developed. All associated south to Vila Verde in the north, Canrightiopsis was isolated stamens in the mesofossil floras including clearly an important component of the Early Cretac- those with pollen similar to that of Canrightiopsis eous vegetation in Portugal. Three species have been have tetrasporangiate anthers and we therefore infer distinguished. Canrightiopsis intermedia is common in tetrasporangiate anthers also for Canrightiopsis. the Famalicão mesofossil flora with about 50 specimens isolated from the organic residue. This species is also recorded from the Buarcos and Vale de Água Comparison of Canrightiopsis with other Early Creta- localities. Canrightiopsis crassitesta is known from the ceous fossils Catefica locality and C. dinisii from the Chicalhão locality. There are also many abraded fruits and iso- Three Early and mid-Cretaceous genera with small lated seeds of Canrightiopsis from these localities as resinous fruits and monocolpate-reticulate pollen well as from two other localities (Arazede and Vila show some similarity with the new fossils described Verde) that have not been assigned at the specific level here: Canrightia from the Early Cretaceous of Portu- due to lack of critical information. Most of them gal (Friis & Pedersen 2011), Zlatkocarpus from the probably belong to one of the three species described mid-Cretaceous of the Czech Republic (Kvaček & here, but there may also be further diversity among Friis 2010), and Couperites from the Early and mid- the small fruits that has not yet been detected. Cur- Cretaceous of eastern North America (Pedersen et al. rently, Canrightiopsis is known only from Portugal and 1991). As indicated by the name, the fossil fruits of has not been recorded in any of the Early Cretaceous Canrightiopsis show similarity with those of Canrightia mesofossil floras from the Potomac group of eastern described from the same plant bearing horizons in North America. Portugal. Fruits of the two genera are small, elliptical

← Figure 11. SRXTM images of Canrightiopsis intermedia gen. et sp. nov. (A–D, G–I) and Canrightiopsis sp. (E, F). Internal structures, from the Early Cretaceous Famalicão locality, Portugal (sample Famalicão 25). A–C. Transverse (A, B) and longitudinal (C) orthoslices through holotype showing fruit and enclosed seed; outer cells of fruit wall collapsed; endotesta (o-en) consists of a single layer of crystal cells with fibrous infilling; larger cells to the outside of the endotesta appears infilled with amorphous substance and may represent exotesta (o-ex? and arrows); internally, there are remnants of tegmen (i-te); subapical position of chalaza (ch) and micropyle (mi) towards the base seen in longitudinal orthoslice; remnants of embryo is preserved in the micropylar part of the seed; secondary crystal infilling (probably pyrite), particularly in endotesta is seen as white flecks (S174033). D. Longitudinal orthoslice through fruit with seed in micropylar area, showing palisade-shaped cells of endotesta (o-en), pointed micropyle (mi) formed from tegmen (i-te) extending through micropylar opening in endotesta and remnants of embryo is preserved in the micropylar part of the seed (em) (S174148). E, F. Transverse (E) and longitudinal (F) orthoslices through fruit with enclosed seed showing cells of endotesta (o-en) and radially elongated cells of tegmen (i-te), perhaps representing an endothelium (S174149). G. Longitudinal orthoslice through fruit and seed showing position of oil cells (asterisk) in the Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 fruit wall; seed wall composed of endotesta (o-en) and tegmen (i-te) and larger cells (arrow) of possible exotesta, hollow space indicating position of embryo (em) and remains of endosperm (e) (S174006). H. Transverse orthoslice through fruit and seed showing endotesta (o-en) filled secondarily with crystals and fragmented tegmen (i-te) (S174004). I. Longitudinal orthoslice showing detail of seed wall with palisade- shaped cells of endotesta (o-en); one of the cells almost completely filled by secondary crystals seen as white flecks (S174005). Scale bars – 500 µm (A, C, E–H), 100 µm (B, D, I).

→ Figure 12. SRXTM images of Canrightiopsis crassitesta gen. et sp. nov. Internal structures, from the Early Cretaceous Catefica locality, Portugal (sample Catefica 49). A–E. Longitudinal (A–D) and transverse (E) orthoslices of fruit in different directions showing thin-walled cells of collapsed fruit wall and thick endotesta (o-en); palisade cells of endosperm with fibrous infilling and scattered crystals (best seen in C); endosperm (e) partly preserved inside the seed and a narrow hollow in the endosperm marks the position of the embryo (em); stigmatic area marked by asterisk and vascular bundle to chalaza (ch) by arrowhead (S174159). F–G. Longitudinal orthoslices of fruit in two directions showing thin-walled cells of collapsed fruit wall, thick endotesta (o-en), and thin- walled cells of tegmen (i-te); apical chalaza (ch)seenin(F) and basal micropyle (mi)seenin(G) (S174039). H. Detail of fruit and seed wall in same specimen as in (F, G), showing the collapsed cells of the fruit and the high endotestal cells (o-en). Scale bars – 500 µm (A, B, E–G), 200 µm (C, D), 100 µm (H). Canrightiopsis, a new Early Cretaceous fossil 203 Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 204 E. M. Friis et al.

to oval or almost spherical berries with ethereal oil monocolpate-reticulate nature of the grains and the cells (or resin bodies) in the fruit wall, and often beaded ornamentation of the muri. However, Couper- wrinkled surface that may obscure details on the ites pollen is distinct from typical Clavatipollenites pol- fruit wall. Fruits of both genera also have distinct len and pollen of extant Chloranthaceae with distal scars from stamens on the fruit wall indicating a apertures (Ascarina and Hedyosmum) in having sharply semi-inferior ovary. Pollen grains observed on the delimited colpus margins, while pollen of Clavatipolle- stigmatic surfaces and fruit wall are monocolpate, nites, Canrightiopsis, Ascarina and Hedyosmum has a semitectate and reticulate. Seeds are pendant, ortho- gradual transition from colpus membrane to the reti- tropous and endotestal with endotesta consisting of a culum of the main body (see also Friis et al. 2011). single layer of palisade cells with crystals and fibrous Although both seed and pollen features suggest a posi- infilling. In other features, Canrightiopsis is clearly tion distinct from Chloranthaceae, a possible chlor- distinct from Canrightia. Most notably, in Canright- anthaceous affinity of Couperites has been addressed iopsis, there are three stamen scars on one side of the in several subsequent phylogenetic analyses (Doyle & ovary in contrast to the radial arrangement of the four Endress 2014, and references cited therein). or five stamens in Canrightia; Canrightiopsis also lacks Clavatipollenites was first described based on dis- a clearly delimited hypanthium, while in Canrightia persed grains from the Early Cretaceous (Barremian– the hypanthium is distinct. Further, Canrightiopsis has Aptian) of southern England (Couper 1958). Origin- one-seeded berries, while Canrightia has two to five ally, a single species, Clavatipollenites hughesii Couper, seeds per fruit, and the beaded supratectal ornamen- was included and studied using light microscopy only. tation of the muri in the Clavatipollenites-type pollen SEM studies of Clavatipollenites-type pollen from the of Canrightiopsis is clearly distinct from the smooth, type horizon by Hughes et al. (1979)demonstrateda unsculptured muri of the Retimonocolpites-type pollen much higher diversity than is discernible in light micro- of Canrightia. scopy (LM) studies, particularly in the supratectal Zlatkocarpus from the Cenomanian of the Bohe- ornamentation. Current practice is, therefore, to use mian Basin includes two species, Z. brnikianus J. the circumscription of the genus indicated by Couper Kvaček et E.M.Friis and Z. pragensis (J.Kvaček et (1960) and, later, by Walker and Walker (1984)for H.Eklund) J.Kvaček et E.M.Friis, both of which monoaperturate, reticulate grains with aperture config- have small resinous fruits comparable to those of uration and supratectal ornamentation closely compar- Canrightiopsis in general morphology, but as in Can- able to that of extant Ascarina (see also discussion in rightia, there is a distinct hypanthium (Kvaček & Friis et al. 2011). Clavatipollenites is very common in Eklund 2003; Kvaček & Friis 2010). The fruits of Early Cretaceous palynofloras with the earliest record Zlatkocarpus are apparently one-seeded, but none of from the Hauterivian of Israel (Brenner & Bickoff the specimens is sufficiently well-preserved to reveal 1992). It is also common in the Early Cretaceous details of seed organisation or internal structures. palynofloras of Portugal represented by several differ- Associated pollen is of the Retimonocolpites-type; ent species (e.g. Pais & Reyre 1981; Trincão 1990; monocolpate and semitectate-reticulate with a long Heimhofer 2004; Heimhofer et al. 2007;Mendes colpus and smooth muri. A recent phylogenetic ana- et al. 2014a). None of the dispersed grains from Por- lysis by Doyle and Endress (2014) resolved Zlatko- tugal are figured at high resolution and detailed com- carpus as closely related to Chloranthaceae, at the parison with the Canrightiopsis pollen is not possible. stem or as member of the crown group. Isolated stamens with Clavatipollenites-type pollen Couperites from the mid-Cretaceous Mauldin Moun- in situ are also common in the Early Cretaceous meso- Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 tain mesofossil flora (Pedersen et al. 1991)issimilarto fossil floras of Portugal. A summary of early angios- Canrightiopsis in having small, one-seeded fruits with perm diversification in Portugal (Friis et al. 1999) ethereal oil cells (or resin bodies) in the fruit wall and listed ten different kinds of monoaperturate, semitec- associated monocolpate, semitectate-reticulate pollen tate-reticulate pollen types with beaded muri, most of with beaded muri. A close relationship of Couperites them are Clavatipollentites-type and a few are Astero- with extant Chloranthaceae was initially suggested by pollis-type. Since then new material has been added. Pedersen et al. (1991), although the authors concluded Based on stamen size and shape, there are clearly that particularly seed characters excluded an assign- several different species (E. M. Friis and K. R. Ped- ment of Couperites to this family. Seed characters also ersen, work in progress), but currently none of the distinguish Couperites from Canrightiopsis. The seeds of isolated stamens can be linked to Canrightiopsis. Couperites are anatropous and exotestal in contrast to the orthotropous and endotestal seeds of Canrightiopsis Comparison with extant Chloranthaceae and extant Chloranthaceae (Ascarina, Chloranthus, Sarcandra). The pollen of Couperites was originally A suite of characters unequivocally links Canrightiopsis referred to as Clavatipollenites-type based on the to members of the Chloranthaceae (Figures 15, 16). Canrightiopsis, a new Early Cretaceous fossil 205 Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016

Figure 13. SRXTM images of Canrightiopsis dinisii gen. et sp. nov. Internal structures, from the Early Cretaceous Chicalhão site, Portugal (holotype, P0311, sample Chicalhão 96). A, B. Transverse orthoslices through fruit with enclosed seed in overview (A) and detail (B); fruit wall of partly collapsed, thin-walled cells; note layer of larger thin-walled cells (arrow) between outer fruit wall and endotesta perhaps representing outer cells of testa; fragmented tegmen (i-te) preserved inside the endotesta. C. Longitudinal orthoslice of fruit showing partly preserved cells of fruit wall toward the apex and section through the stigma (asterisk); seed wall consists of endotesta (o-en) and fragmented tegmen (i-te) and perhaps outer testa (arrow). D. Tangential and longitudinal orthoslice through seed showing the foveolate-grooved surface of endotesta. Scale bars – 500 µm (A, C), 250 µm (D), 100 µm (B). 206 E. M. Friis et al.

while pollen of Chloranthus and Sarcandra typically has smooth muri. The Clavatipollenites-type pollen of Canrightiopsis is particularly similar to that of extant Ascarina. Resemblance of isolated Clavatipollenites to Ascarina was already pointed out earlier by Couper (1960) and Walker and Walker (1984). Pollen of Ascarina is the only extant Chloranthaceae pollen with a distal colpus that is simple or sometimes three-armed. Pollen of Hedyosmum also has a single distal aperture, but the aperture is diffusely five- armed. Pollen of Sarcandra is polyporate and that of Chloranthus variable, spiraperturate to polycolpate. None of them has distal apertures. A distal aperture is, however, probably present in the peculiar pollen of Chloranthistemon alatus H. Eklund, K.R.Pedersen et E. M. Friis. As in Canrightiopsis, fruits of Ascarina, Chloranthus and Sarcandra are berries with a single orthotropous, pendant and endotestal seed. The testa in Canright- iopsis may consists of only two layers of cells, an outer layer of large, thin-walled cells (exotestal) and an inner layer (endotesta) of palisade-shaped cells with fibrous infilling and scattered crystals. The same condition is known for Ascarina, while the Figure 14. Schematic outline of ovule structure of Canrightiopsis testa in Chloranthus and Sarcandra consists of several from the Early Cretaceous of Portugal. Fruit wall is indicated by layers of cells (Endress & Igersheim 1997). Because light green colour and oil cells in fruit wall not coloured. The the delimitation of the outer testa and fruit wall is not outer integument (testa) is indicated in light yellow and separated in an outer layer of thin walled cells perhaps representing exotesta distinct in the fossil material, we have not included (o-ex?) and an inner endotestal layer with crystals (o-en). The inner this character in the analysis. integument (tegmen, i-te) is indicated by a brownish colour; note Ethereal oil cells are present in the surface of the that the structure of tegmen is not fully understood. fruit wall of Canrightiopsis as well as in Chloranthus and Sarcandra, while in Ascarina the oil cells are deeper in the fruit wall (Endress & Igersheim Of particular importance is the unusual androecium in 1999). The embryo in mature seeds is small sur- the two bisexual members of Chloranthaceae (Chlor- rounded by copious endosperm in Canrightiopsis anthus and Sarcandra)andinCanrightiopsis with sta- and in all extant Chloranthaceae. mens borne on one side of the ovary, one to two thirds up on the ovary, without preceding a perianth or dis- Canrightiopsis, an evolutionary link between Canrigh- tinguishable hypanthium (Endress 1987). A similar tia and Chloranthus–Sarcandra? arrangement is not known for any other extant angiosperm. Based on the protruding scars on the fruit wall In the character analysis, Canrightia differs from the Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 Canrightiopsis has three stamens: one borne centrally reconstructed common ancestor of extant Chlor- and two borne laterally with one on each side of the anthaceae only by the number of carpels and ovules central stamen, about 300 µm from the central stamen. (Figure 17) and it may represent a plesiomorphic In Chloranthus, the androecium is tripartite typically ancestral Chloranthaceae (see also Doyle & Endress with a broad base bearing a central, dithecate and 2014), while Zlatkocarpus may represent an extinct tetrasporangiate lobe and two lateral, monothecate side lineage, not directly related to any of the mod- and bisporangiate lobes with the sporangia of the lat- ern Chloranthaceae. The analysis further shows close eral lobes borne on the outer margin (facing away from resemblance between the character suites of Can- the central lobe). In Sarcandra,thereisasingle,cen- rightiopsis and the hypothetical common ancestors tral, dithecate and tetrasporangiate, stamen. As in of Chloranthus–Sarcandra and Ascarina–Chloranthus– extant Chloranthaceae, the subtending bracts are not Sarcandra. This configuration suggests an evolution- attached to the fruits. ary pathway in the Chloranthaceae clade that links Pollen in all extant members of the Chloranthaceae Canrightia and Canrightiopsis to the extant members is semitectate-reticulate with beaded muri in Ascar- of the Chloranthaceae and provides a simple expla- ina, Hedyosmum and occasionally in Chloranthus, nation for the derivation of the unusual floral Canrightiopsis, a new Early Cretaceous fossil 207

Figure 15. Phylogenetic position of Canrightia, Canrightiopsis and Zlatkocarpus based on a data set of 63 morphological characters. Shown is a planar phylogenetic network, a neighbour-net splits graph, inferred from pairwise (Hamming) distances calculated from the character matrix. Members of commonly accepted clades are generally grouped and the reduced character set is sufﬁcient to group extant Chloranthaceae except for Hedyosmum in the neighbour-net, with the three fossil taxa resolved as close relatives of the core Chloranthaceae (Ascarina, Chloranthus, Sacandra). Canrightiopsis is resolved as most similar to extant Chloranthus and Sarcandra in its preserved morphological traits.

‘ ’ Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 Figure 16. Balls-and-sticks graph with proportional branch lengths depicting the result of the ancestral state reconstruction under maximum likelihood. Ancestral states were reconstructed for 11 characters that differ among extant members of the Chloranthaceae. States of two characters (character 7, 13), iden- tical among extant members, are shown for comparison with the fossil taxa. Each ‘ball’ shows the relative probability of a character state at the given node. Char- acter states observed in fossil members of Chloranthaceae are shown to the right. The branch (‘stick’)lengthsreﬂect the amount of molecular evolution (expected substitutions per site). 208 E. M. Friis et al.

Figure 17. Trait evolution in Chlor- anthaceae. The synopsis is based on reconstructed states for hypothetical ancestors of extant genera (cf. Figure 16) and empirical evidence from fossil members of the lineage. The only con- ﬂict between reconstruction and empirical evidence is highlighted by red colouring: According to the ancestral state reconstruction, tepals would have been already reduced in the common ancestor of all extant taxa. Hedyosmun would then have de-evolved to an ancestral state (reversal), which is unlikely. In contrast, the situation in Canrightiopsis indicates that tepals were subsequently reduced in the stem lineage of core Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 Chloranthaceae Ascarina, Chloranthus and Sarcandra.

structure and androecium of Chloranthus and Sar- compared to the other three branches in the mole- candra (Figure 18). cular-based trees (Figure 16). Extant Hedyosmum deviates markedly from Can- Under the currently accepted rooting scenario rightia and the common ancestor of all other Chlor- (Qiu et al. 1999;Zhang&Renner2003;Doyle& anthaceae in having unisexual flowers with Endress 2014), loss of tepals, reduction of stamen unistaminate male flowers, pollen grains with irregu- number and displacement of the androecium to one lar aperture, epigynous pistillate flowers with well- side would represent putative synapomorphies (or developed style, lack of oil cells in the fruit wall, dry more precise: shared apomorphies) of the lineage fruit and unspecialised endotesta. The derived posi- leading to Ascarina, Chloranthus and Sarcandra. tion of Hedyosmum is also corroborated by the much Thus, Canrightiopsis may be considered as an longer root to tip distance of its terminal branch early, yet relatively plesiomorphic representative of Canrightiopsis, a new Early Cretaceous fossil 209

Figure 18. Proposed evolutionary sequence from the bisexual, radial symmetrical and partly epigynous ﬂowers of Canrightia to the bisexual, monosymmetrical and naked ﬂowers of Canrightiopsis, Chloranthus and Sarcandra, illustrating also the three-stamen scenario for the derivation of the tripartite androecium of Chloranthus.

Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 the crown group of Chloranthaceae, probably pre- suggesting that the hypanthium is completely dating the split between Ascarina and Chloranthus– absorbed by the ovary wall in Canrightiopsis, Chlor- Sarcandra. Under this model, the change from one anthus and Sarcandra. The androecium of Can- to many (non-distal) apertures is a shared apomor- rightiopsis has a reduced number of stamens phy of Chloranthus–Sarcandra. Ascarina is distin- compared to that of Canrightia that could have guished from the common ancestor/Canrightiopsis developed by loss of one or two ventral stamens by its unisexual ﬂowers and apparent superior and a displacement of the remaining three stamens ovary. to one side. Only few evolutionary steps further are In Canrightia and Zlatkocarpus, the perianth is required for deriving the tripartite (three-lobed) strongly reduced, represented by short tips on the androecium of Chloranthus and the unistaminate rim of the hypanthium. In Canrightiopsis as well as androecium of Sarcandra from the three stamens in extant Ascarina, Chloranthus and Sarcandra,the of Canrightiopsis: further displacement of the two perianth parts are completely lost and there is no lateral stamens towards the centre; concrescence of obvious hypanthium. However, stamens/stamen the three stamen bases; loss of one set of sporangia scars are borne at the same level as in Canrightia from the lateral stamens in Chloranthus;andlossof 210 E. M. Friis et al.

both lateral stamens in Sarcandra. Concrescence of Acknowledgements stamens could have happened before or after loss The authors thank Marco Stampanoni, Federica of sporangia. The latter scenario is in accordance Marone (PSI) and Anna Lindström (NRM) for help with one of the interpretation of the Chloranthus with the SRXTM analyses performed at the Swiss androecium presented by Swamy (1953). In both Light Source, Paul Scherrer Institute (PSI), Villigen, extant Chloranthus and in fossil Chloranthistemon, Switzerland. Pollyanna von Knorring (NRM) is there are species in which the basal parts of the thanked for help with the illustration in Figure 17. individual lobes are free. The three-stamen model The authors also thank Peter K. Endress, University contrasts the single-stamen models by Endress of Zürich, Switzerland, and Patrick Herendeen, Chi- (1987). In these models, the tripartite androecium cago Botanical Garden, Chicago, U.S.A., for helpful is derived from a single stamen either by polymer- comments and suggestions. Financial support by the isation, concrescence and reduction or by lobation Swiss Light Source (European Union FP6 projects and fractionation. In both single stamen scenarios, 20110963 and 20130185), by the Swedish Research thethecaeofthelaterallobesareborneonthe Council (621-2011-5431) and by CretaCarbo project outer margin. In the Chloranthistemon-like fossils (PTDC/CTE-GIX/113983/2009) is gratefully from mid-Cretaceous of Germany, the thecae of acknowledged. the lateral lobes appear to be borne on the inner margins (Friis et al. 2011; Hartkopf-Fröder et al. 2011), which may also have implications for Supplemental data understanding the derivation of the tripartite androecium. However, the apparent diverging pat- Supplemental data for this article can be accessed tern in the German taxon may be an artefact here. caused by compression (Peter K. Endress, perso- nal communication, April 2015), and further studies are needed to establish the nature of this mid- Disclosure statement Cretaceous fossil. No potential conflict of interest was reported by the authors. Conclusion The new fossil genus, Canrightiopsis, is important in References several respects. It represents a new extinct angiosperm and further documents the extensive diversity Angiosperm Phylogeny Group (APG). 2009. An update of the Angiosperm Phylogeny Group classification for the orders among basal angiosperms in the Early Cretaceous fl fl fi and families of owering plants: APG III. Botanical Journal mesofossil oras. It is also signi cant in corroborat- of the Linnean Society 161: 105–121. doi:10.1111/(ISSN) ing the importance of Chloranthaceae in early 1095-8339. angiosperm diversification and provides a key for Brenner GJ, Bickoff IS. 1992. Palynology and age of the Lower interpreting the unusual floral structure and androe- Cretaceous basal Kurnub group from the coastal plain to the cium in extant Chloranthus and Sarcandra. An evolu- northern Negev of Israel. Palynology 16: 137–185. doi:10.1080/ 01916122.1992.9989411. tionary sequence from the bisexual, radial Bryant D, Moulton V. 2002. NeighborNet: An agglomerative symmetrical and partly epigynous flowers of Can- method for the construction of planar phylogenetic networks. rightia to the bisexual, monosymmetrical and naked In: Guigó R, Gusfield O, eds. Second Workshop on Bioinfor- Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 flowers of Canrightiopsis, Chloranthus and Sarcandra matics (WABI) 2002. LNCS 2452, 375–391. Berlin, Heidel- is proposed. Based on the new fossils, we also pro- berg: Springer. Bryant D, Moulton V. 2004. NeighborNet: An agglomerative pose that the tripartite androecium of Chloranthus is method for the construction of phylogenetic networks. Molecu- derived from closing up of three full stamens towards lar Biology and Evolution 21: 255–265. doi:10.1093/molbev/ the centre of the dorsal side followed by concres- msh018. cence and loss of sporangia, contrasting the prevail- Couper RA. 1958. British Mesozoic microspores and pollen ing single stamen hypotheses where the tripartite grains. A systematic and stratigraphic study. Palaeontographica Abteilung B 103: 75–179. androecium is thought to have developed from a Couper RA. 1960. New Zealand Mesozoic and Cainozoic plant single stamen. Canrightiopsis occurs abundantly in microfossils. New Zealand Geological Survey. Palaeontological the Portuguese mesofossil floras from the Early Cre- Bulletin 32: 1–87. taceous and was clearly an important element in the Crane PR, Friis EM, Pedersen KR. 1989. Reproductive structure Early Cretaceous vegetation of the region. So far, it and function in Cretaceous Chloranthaceae. Plant Systematics and Evolution 165: 211–226. doi:10.1007/BF00936003. has not been recorded from the more or less con- Denk T, Grimm GW. 2009. The biogeographic history of beech temporary mesofossil floras of eastern North trees. Review of Palaeobotany and Palynology 158: 83–100. America. doi:10.1016/j.revpalbo.2009.08.007. Canrightiopsis, a new Early Cretaceous fossil 211

Dinis JL 2001. Definição da Formação da Figueira da Foz - Botanical Journal of the Linnean Society 172: 465–477. Aptiano a Cenomaniano do sector central da margem oeste doi:10.1111/boj.12065. ibérica /Definition of the Figueira da Foz Formation - Aptian Hartkopf-Fröder C, Rust J, Wappler T, Friis EM, Viehofen A. to Cenomanian of the central sector of the western Iberian 2011. Mid-Cretaceous charred fossil flowers reveal direct margin. Comunicações do Instituto Geológico e Mineiro 88: observation of arthropod feeding strategies. Biological Letters 127–160. doi:10.1098/rsbl.2011.0696. Dinis JL, Rey J, Cunha PP, Callapez P, Pena dos Reis R. 2008. Hedlund RW, Norris G. 1968. Spores and pollen grains from Stratigraphy and allogenic controls of the western Portugal Fredericksburgian (Albian) strata, Marshall County, Okla- Cretaceous: An updated synthesis. Cretaceous Research 29: homa. Pollen et Spores 10: 129–159. 772–780. doi:10.1016/j.cretres.2008.05.027. Heimhofer U. 2004. Response of terrestrial palaeoenvironemnts to Doyle JA, Endress PK. 2010. Integrating Early Cretaceous fossils past changes in climate and carbon-cycling: Insights from into the phylogeny of living angiosperms: Magnoliidae and palynology and stable isotope geochemistry. PhD Thesis, eudicots. Journal of Systematics and Evolution 48: 1–35. ETH, Zurich, Switzerland. doi:10.1111/jse.2010.48.issue-1. Heimhofer U, Hochuli PA, Burla S, Weissert H. 2007. New Doyle JA, Endress PK. 2014. Integrating Early Cretaceous fossils records of Early Cretaceous angiosperm pollen from Portu- into the phylogeny of living angiosperms: ANITA lines and guese coastal deposits: Implications for the timing of the early relatives of Chloranthaceae. International Journal of Plant angiosperm radiation. Review of Palaeobotany and Palynology Sciences 175: 555–600. doi:10.1086/675935. 144: 39–76. doi:10.1016/j.revpalbo.2005.09.006. Eklund H, Friis EM, Pedersen KR. 1997. Chloranthaceous floral Herendeen PS, Crepet WL, Nixon KC. 1993. Chloranthus-like structures from the Late Cretaceous of Sweden. Plant Systematics stamens from the Upper Cretaceous of New Jersey. American and Evolution 207: 13–42. doi:10.1007/BF00985207. Journal of Botany 80: 865–871. Endress PK. 1987. The Chloranthaceae: Reproductive structures Hintermüller C, Marone F, Isenegger A, Stampanoni M. 2010. and phylogenetic position. Botanische Jahrbücher für Systema- Image processing pipeline for synchrotron-radiation-based tik. Pflanzengeschichte und Pflanzengeographie 109: 153–226. tomographic microscopy. Journal of Synchrotron Radiation Endress PK, Igersheim A. 1997. Gynoecium diversity and sys- 17: 550–559. doi:10.1107/S0909049510011830. tematics of the Laurales. Botanical Journal of the Linnean Hughes NF. 1994. The enigma of angiosperm origins. Cambridge: Society 125: 93–168. doi:10.1111/boj.1997.125.issue-2. Cambridge University Press. Endress PK, Igersheim A. 1999. Gynoecium diversity and sys- Hughes NF, Drewry G, Laing JF. 1979. Barremian earliest tematics of the basal eudicots. Botanical Journal of the Linnean angiosperm pollen. Palaeontology 22: 513–536. Society 130: 305–393. doi:10.1111/boj.1999.130.issue-4. Huson DH, Bryant D. 2006. Application of phylogenetic networks Friis EM, Crane PR, Pedersen KR. 1997. Fossil history of mag- in evolutionary studies. Molecular Biology and Evolution 23: noliid angiosperms. In: Iwatsuki K, Raven PH, eds. Evolution 254–267. doi:10.1093/molbev/msj030. and diversification of land plants, 121–156. Tokyo, Berlin, Katoh K, Standley DM. 2013. MAFFT multiple sequence align- Heidelberg, New York: Springer-Verlag. ment software version 7: Improvements in performance and Friis EM, Crane PR, Pedersen KR. 2011. Early flowers and angios- usability. Molecular Biology and Evolution 30: 772–780. perm evolution. Cambridge: Cambridge University Press. doi:10.1093/molbev/mst010. Friis EM, Marone F, Pedersen KR, Crane PR, Stampanoni M. Kvaček J, Eklund H. 2003. A report on newly recovered repro- 2014. Three-dimensional visualization of fossil flowers, fruits, ductive structures from the Cenomanian of Bohemia (Central seeds, and other plant remains using synchrotron radiation X- Europe). International Journal of Plant Sciences 164: 1021– ray tomographic microscopy (SRXTM): New insights into 1039. doi:10.1086/ijps.2003.164.issue-6. Cretaceous plant diversity. Journal of Paleontology 88: 684– Kvaček J, Friis EM. 2010. Zlatkocarpus gen. nov., a new angiosperm 701. doi:10.1666/13-099. reproductive structure with monocolpate-reticulate pollen from Friis EM, Pedersen KR. 2011. Canrightia resinifera gen. et sp. the Late Cretaceous (Cenomanian) of the Czech Republic. nov., a new extinct angiosperm with Retimonocolpites-type pol- Grana 49: 115–127. doi:10.1080/00173134.2010.481845. len from the Early Cretaceous of Portugal: Missing link in the Lewis PO. 2001. A likelihood approach to estimating phylogeny eumagnoliid tree? Grana 50: 3–29. doi:10.1080/00173134. from discrete morphological character data. Systematic Biol- 2011.559728. ogy 50: 913–925. doi:10.1080/106351501753462876. Friis EM, Pedersen KR, Crane PR. 1999. Early angiosperm diver- Maddison WP, Maddison DR. 2011. Mesquite: A modular sys- sification: The diversity of pollen associated with angiosperm tem for evolutionary analysis. Version 2.75. http://mesquitepro Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016 reproductive structures in Early Cretaceous floras from Portu- ject.org. gal. Annals of the Missouri Botanical Garden 86: 259–296. Martínez C, Madriñán S, Zavada M, Alberto Jaramillo C. 2013. Friis EM, Pedersen KR, von Balthazar M, Grimm GW, Crane PR. Tracing the fossil pollen record of Hedyosmum (Chloranthaceae), 2009. Monetianthus mirus gen. et sp. nov., a nymphaealean an old lineage with recent Neotropical diversification. Grana 52: flower from the Early Cretaceous of Portugal. International 161–180. doi:10.1080/00173134.2012.760646. Journal of Plant Sciences 170: 1086–1101. doi:10.1086/592013. Mendes MM, Dinis J, Pais J, Friis EM. 2014a. Vegetational Göker M, García-Blázquez G, Voglmayr H, Tellería MT, Martín composition of the Early Cretaceous Chicalhão flora (Lusita- MP. 2009. Molecular taxonomy of phytopathogenic fungi: A nian Basin, western Portugal) based on palynological and case study in Peronospora. PLoS ONE 4: e6319. mesofossil assemblages. Review of Palaeobotany and Palynol- Göker M, Grimm GW. 2008. General functions to transform ogy 200: 65–81. doi:10.1016/j.revpalbo.2013.08.003. associate data to host data, and their use in phylogenetic infer- Mendes MM, Grimm GW, Pais J, Friis EM. 2014b. Fossil ence from sequences with intra-individual variability. BMC Kajanthus lusitanicus gen. et sp. nov. from Portugal: Floral Evolutionary Biology 8: 86. evidence for Early Cretaceous Lardizabalaceae (Ranunculales, Grimm GW, Renner SS. 2013. Harvesting GenBank for a Betu- basal eudicot). Grana 53: 283–301. doi:10.1080/00173134. laceae supermatrix, and a new chronogram for the family. 2014.932431. 212 E. M. Friis et al.

Nichols DJ, Matsukawa M, Ito M. 2006. Palynology and age of Stamatakis A, Hoover P, Rougemont J. 2008. A rapid bootstrap some Cretaceous non-marine deposits in Mongolia and China. algorithm for the RAxML web servers. Systematic Biology 57: Cretaceous Research 27: 241–251. doi:10.1016/j.cretres.2005. 758–771. doi:10.1080/10635150802429642. 11.004. Stampanoni M, Groso A, Isenegger A, Mikuljan G, Chen Q, Pais J, Reyre Y. 1981. Problèmes posés par la population sporo- Bertrand A, Henein S, Betemps R, Frommherz U, Bohler P, pollinique d’un niveau à plantes de la série de Buarcos (Portu- Meister D, Lange M, Abela R. 2006. Trends in synchrotron- gal). Boletim da Sociedade Geológica de Portugal 22: 35–40. based tomographic imaging: The SLS experience. In: Bonse Pattengale ND, Masoud A, Bininda-Emonds ORP, Moret BME, U, ed. Developments in X-ray tomography V, Vol. 6318. San Stamatakis A. 2009. How many bootstrap replicates are neces- Diego: Proceedings of SPIE-The International Society for sary? In: Batzoglou S, ed. RECOMB 2009, 184–200. Berlin, Optical Engineering. Heidelberg: Springer. Swamy BGL. 1953. The morphology and relationships of the Pedersen KR, Crane PR, Drinnan AN, Friis EM. 1991. Fruits Chloranthaceae. Journal of the Arnold Arboretum 34: 375–411. from the mid-Cretaceous of North America with pollen grains Swofford DL. 2002. PAUP*: Phylogenetic analysis using parsi- of the Clavatipollenites type. Grana 30: 577–590. doi:10.1080/ mony (* and other methods). Champaign: National Illinois 00173139109427816. History Survey. Qiu Y-L, Lee JH, Bernasconi-Quadroni F, Soltis DE, Soltis PS, Zanis Trincão PRP. 1990. Esporos e pólenes do Cretácio inferior (Ber- M, Zimmer EA, Chen ZD, Savolainen V, Chase MW. 1999. The riasiano-Aptiano) de Portugal: Paleontologia e biostratigraﬁa. earliest angiosperms: Evidence from mitochondrial, plastid and PhD Thesis, Universidade Nova de Lisboa, Lisbon. nuclear genomes. Nature 402: 404–407. doi:10.1038/46536 Walker JW, Walker AG. 1984. Ultrastructure of Lower Cretac- Rey J, Dinis JL, Callapez P, Cunha PP. 2006. Da rotura conti- eous angiosperm pollen and the origin and early evolution of nental à margem passiva. Composição e evolução do Cretácico ﬂowering plants. Annals of the Missouri Botanical Garden 71: de Portugal. Lisbon: Ministério da Economia e da Inovação. 464–521. Stamatakis A. 2006. RAxML-VI-HPC: Maximum-Likelihood- Zhang L-B, Renner SS. 2003. The deepest splits in chloranthaceae based phylogenetic analyses with thousands of taxa and as resolved by chloroplast sequences. International Journal of mixed models. Bioinformatics 22: 2688–2690. doi:10.1093/ Plant Sciences 164: S383–S392. doi:10.1086/ijps.2003.164. bioinformatics/btl446. issue-s5. Downloaded by [Statsbiblioteket Tidsskriftafdeling] at 04:18 18 January 2016